Hyperbranched poly(β-amino ester) based polyplex nanopaticles for delivery of CRISPR/Cas9 system and treatment of HPV infection associated cervical cancer

Xueqin Gao; Zhuang Jin; Xiangyu Tan; Chong Zhang; Chenming Zou; Wei Zhang; Jiahui Ding; Bhudev C Das; Konstantin Severinov; Inga Isabel Hitzeroth; Priya Ranjan Debata; Dan He; Xin Ma; Xun Tian; Qinglei Gao; Jun Wu; Rui Tian; Zifeng Cui; Weiwen Fan; Zhaoyue Huang; Chen Cao; Yuxian Bao; Songwei Tan; Zheng Hu

doi:10.1016/j.jconrel.2020.02.045

Hyperbranched poly(β-amino ester) based polyplex nanopaticles for delivery of CRISPR/Cas9 system and treatment of HPV infection associated cervical cancer

J Control Release. 2020 May 10:321:654-668. doi: 10.1016/j.jconrel.2020.02.045. Epub 2020 Feb 28.

Authors

Xueqin Gao¹, Zhuang Jin², Xiangyu Tan³, Chong Zhang⁴, Chenming Zou¹, Wei Zhang⁴, Jiahui Ding⁴, Bhudev C Das⁵, Konstantin Severinov⁶, Inga Isabel Hitzeroth⁷, Priya Ranjan Debata⁸, Dan He⁹, Xin Ma¹⁰, Xun Tian¹¹, Qinglei Gao³, Jun Wu¹², Rui Tian², Zifeng Cui², Weiwen Fan², Zhaoyue Huang², Chen Cao¹¹, Yuxian Bao¹³, Songwei Tan¹⁴, Zheng Hu¹⁵

Affiliations

¹ Department of Pharmacy, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
² Department of Obstetrics and Gynecology, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China.
³ Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei, China.
⁴ School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
⁵ Amity Institute of Molecular Medicine & Stem Cell Research, Amity University, Uttar Pradesh, Noida 201313, India.
⁶ Skolkovo Institute of Science and Technology, Skolkovo, Moscow Region 143025, Russia.
⁷ Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, South Africa.
⁸ Department of Zoology, North Orissa University, Takatpur, Baripada, Odisha 757003, India.
⁹ Department of Neurology, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, Guangdong, China.
¹⁰ Department of Urology, General Hospital of People's Liberation Army, Beijing 100039, China.
¹¹ Department of Obstetrics and Gynecology, Academician expert workstation, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, Hubei, China.
¹² School of Biomedical Engineering, Sun Yat-sen University, Guangzhou 510006, Guangdong, China.
¹³ Generulor Company Bio-X Lab, Guangzhou 510006, Guangdong, China.
¹⁴ School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China. Electronic address: [email protected].
¹⁵ Department of Obstetrics and Gynecology, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China; Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei, China; Precision Medicine Institute, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China. Electronic address: [email protected].

PMID: 32114092
DOI: 10.1016/j.jconrel.2020.02.045

Abstract

Persistent high-risk HPV infection is the main factor for cervical cancer. HPV E7 oncogene plays an important role in HPV carcinogenesis. Down-regulation of E7 oncogene expression could induce growth inhibition in HPV-positive cells and thus treats HPV related cervical cancer. Here we developed a non-virus gene vector based on poly(amide-amine)-poly(β-amino ester) hyperbranched copolymer (hPPC) for the delivery of CRISPR/Cas9 system to specifically cleave HPV E7 oncogene in HPV-positive cervical cancer cells. The diameter of polyplex nanoparticles (NPs) formed by hPPCs/linear poly(β-amino ester) (PBAE) and plasmids were approximately 300 nm. These hPPCs/PBAE-green fluorescence protein plasmids polyplex NPs showed high transfection efficiency and low toxicity in cells and mouse organs. By cleaving HPV16 E7 oncogene, reducing the expression of HPV16 E7 protein and increasing intracellular retinoblastoma 1 (RB1) amount, hPPCs/PBAE-CRISPR/Cas9 therapeutic plasmids polyplex NPs, especially highly branched hPPC1-plasmids polyplex NPs, exhibited strong growth inhibition of cervical cancer cells in vitro and xenograft tumors in nude mice. Together, the hPPCs/PBAE polyplex NPs to deliver HPV16 E7 targeted CRISPR/Cas9 system in this study could potentially be applied to treat HPV-related cervical cancer.

Keywords: CRISPR/Cas9; Cervical cancer; HPV; Hyperbranched copolymer; Poly(β-amino ester).

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
CRISPR-Cas Systems
Drug Delivery Systems
Esters
Female
Humans
Mice
Mice, Nude
Papillomavirus E7 Proteins / genetics
Papillomavirus Infections* / therapy
Polymers*
Uterine Cervical Neoplasms* / genetics
Uterine Cervical Neoplasms* / therapy

Substances

Esters
Papillomavirus E7 Proteins
Polymers
poly(beta-amino ester)